An Intact DNA-binding Domain Is Not Required for Peroxisome Proliferator-activated Receptor γ (PPARγ) Binding and Activation on Some PPAR Response Elements

Peroxisome proliferator-activated receptor γ (PPARγ) interacts with retinoid X receptor (RXR) on PPAR response elements (PPREs) to regulate transcription of PPAR-responsive genes. To investigate the binding of PPARγ and RXR to PPREs, three mutations were constructed in the DNA-binding domains of PPA...

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Veröffentlicht in:The Journal of biological chemistry 2005-02, Vol.280 (5), p.3529-3540
Hauptverfasser: Temple, Karla A., Cohen, Ronald N., Wondisford, Sarah R., Yu, Christine, Deplewski, Dianne, Wondisford, Fredric E.
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container_end_page 3540
container_issue 5
container_start_page 3529
container_title The Journal of biological chemistry
container_volume 280
creator Temple, Karla A.
Cohen, Ronald N.
Wondisford, Sarah R.
Yu, Christine
Deplewski, Dianne
Wondisford, Fredric E.
description Peroxisome proliferator-activated receptor γ (PPARγ) interacts with retinoid X receptor (RXR) on PPAR response elements (PPREs) to regulate transcription of PPAR-responsive genes. To investigate the binding of PPARγ and RXR to PPREs, three mutations were constructed in the DNA-binding domains of PPARγ; two of the mutants maintained the structure of zinc finger I (PPARγ-GS and PPARγ-AA), and a third mutation disrupted the protein structure of zinc finger I (PPARγ-CS). Results indicated that the mutations of PPARγ that maintained intact zinc fingers were capable of binding to a variety of PPREs in the presence of RXR and could activate transcription on several PPREs. In parallel, a mutation was created in the DNA-binding domain of RXRα that maintained the structure of the zinc fingers (RXR-GS) but did not bind DNA and was transcriptionally inactive. Examination of the 3′ half-site of several PPREs revealed that variations from the consensus sequence reduced or abolished transcriptional activity, but conversion to consensus improved transcriptional activity with PPARγ-GS and PPARγ-AA. Examination of the 5′ half-site indicated that the upstream three nucleotides were more important for transcriptional activity than the downstream three nucleotides. Our data demonstrated that stringent binding of RXR to the 3′ half-site of a PPRE is more influential on the binding of the PPARγ/RXR heterodimer than the ability of PPARγ to bind DNA. Thus, unlike RXR, PPARγ exhibits promiscuity in binding on a PPRE, suggesting that the definition of a PPRE for PPARγ may need to be expanded.
doi_str_mv 10.1074/jbc.M411422200
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Examination of the 5′ half-site indicated that the upstream three nucleotides were more important for transcriptional activity than the downstream three nucleotides. Our data demonstrated that stringent binding of RXR to the 3′ half-site of a PPRE is more influential on the binding of the PPARγ/RXR heterodimer than the ability of PPARγ to bind DNA. 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subjects Acyl-CoA Oxidase - genetics
Amino Acid Sequence
Animals
Binding Sites
Consensus Sequence
Dimerization
Mice
Molecular Sequence Data
Mutagenesis
Phosphoenolpyruvate Carboxykinase (GTP) - genetics
PPAR gamma - chemistry
PPAR gamma - genetics
PPAR gamma - metabolism
Protein Binding
Protein Structure, Tertiary
Response Elements - physiology
Retinoid X Receptors - metabolism
Transcriptional Activation - physiology
Transfection
title An Intact DNA-binding Domain Is Not Required for Peroxisome Proliferator-activated Receptor γ (PPARγ) Binding and Activation on Some PPAR Response Elements
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